US9526728B2 - Medical use of a DPP-4 inhibitor - Google Patents

Abstract

The present invention relates to the use of 1-[(4-methyl-quinazolin-2-yl)methyl]-3 -methyl-7-(2-butyn-1-yl)-8-(3-(R)-amino-piperidin-1-yl)-xanthine for use in a patient being at risk of or having heart failure with preserved ejection fraction (HFpEF).

Description

FIELD OF THE INVENTION

The present invention relates to certain DPP-4 inhibitors (preferably linagliptin), optionally in combination with one or more other active agents, for use in a patient (particularly human patient) being at risk of or having heart failure with preserved ejection fraction (HFpEF), particularly in patients with chronic kidney disease (CKD) such as advancing or severe renal impairment or end-stage renal disease (ESRD).

The present invention further relates to certain DPP-4 inhibitors (preferably linagliptin), optionally in combination with other active agents, for use in treating, preventing, reducing the risk of, slowing progression of, delaying the onset of, and/or protecting against heart failure with preserved ejection fraction (HFpEF), particularly in patients (particularly human patients) with chronic kidney disease (CKD) such as advancing or severe renal impairment or end-stage renal disease (ESRD), to pharmaceutical compositions or combinations comprising such active components, and to certain therapeutic uses thereof.

BACKGROUND OF THE INVENTION

Heart failure (HF) is the leading cause of cardiovascular morbidity and mortality worldwide. About half of heart failure patients have heart failure with preserved ejection fraction (HFpEF). Distinct from traditional HF, i.e., heart failure with reduced ejection fraction in which the ventricle cannot contract, patients with HFpEF show declined performance of heart ventricle, not at the time of contraction, but during the phase of diastole. HFpEF patients show normal ejection fraction of blood pumped out of the ventricle, but the heart muscle does not quickly relax to allow efficient filling of blood returning from the body. Morbidity and mortality of HFpEF are similar to traditional HF; however, therapies that benefit traditional HF are less effective in treating or preventing HFpEF.

One of the causes of diastolic dysfunction (diastolic heart failure) can be cardiac fibrosis.

For people with chronic kidney disease (CKD) and advancing severe renal impairment or ESRD, salvage of the native kidney is not a tangible goal with treatment options limited to renal transplantation or dialysis. However, these individuals are also at markedly increased risk of other comorbidities that themselves may remain amenable to therapeutic interventions. Among these comorbidities, heart failure, especially heart failure with preserved ejection fraction (HFpEF) is particularly common, where it carries a mortality risk equivalent to that of systolic heart failure, but has no evidence-based treatment.

HFpEF may affect up to 70% of the dialysis population. Unfortunately, whereas ACE inhibitors and angiotensin II receptor blockers represent established evidence-based therapies for the treatment of either systolic heart failure or CKD they provide little if any benefit in the treatment of HFpEF.

SUMMARY OF THE INVENTION

The present invention relates to a certain DPP-4 inhibitor (preferably linagliptin), optionally in combination with one or more other active agents, for use in a patient (particularly human patient) being at risk of or having heart failure with preserved ejection fraction (HFpEF), particularly in a patient having chronic kidney disease (CKD) such as advancing or severe renal impairment or end-stage renal disease (ESRD), such patient may be diabetic or non-diabetic.

The present invention further relates to a certain DPP-4 inhibitor (preferably linagliptin), optionally in combination with one or more other active agents, for use in treating, preventing, reducing the risk of, slowing progression of, delaying the onset of, and/or protecting against heart failure with preserved ejection fraction (HFpEF) and/or diseases associated therewith (such as e.g. skeletal muscle and vascular dysfunction, (pulmonary) hypertension, renal failure, anaemia, and/or atrial fibrillation; or major adverse cardiovascular events (MACE)), particularly in patients with chronic kidney disease (CKD) such as advancing or severe renal impairment or, particularly, end-stage renal disease (ESRD), such patient may be diabetic or non-diabetic.

Major adverse cardiovascular events (MACE) may include myocardial infarction, stroke, cardiovascular death, and/or cardiovascular hospitalization such as for (unstable or stable) angina pectoris, heart failure or coronary revascularization.

The present invention further relates to a certain DPP-4 inhibitor (preferably linagliptin), optionally in combination with one or more other active agents, for use in improving cardiac or diastolic function and/or reducing cardiac fibrosis or hypertrophy in such patients.

The effects of the DPP-4 inhibitor on HFpEF (e.g. improvement in diastolic function, cardiac fibrosis, etc.) according to the present invention may be beyond or independent from glycemic control and/or beyond or independent from reno-protective effects (renal function or structure).

The present invention further relates to a certain DPP-4 inhibitor (preferably linagliptin), optionally in combination with one or more other active agents, for use in patients with chronic kidney disease (CKD) such as advancing or severe renal impairment or, particularly, end-stage renal disease (ESRD), having heart failure with preserved ejection fraction (HFpEF).

The present invention further relates to a certain DPP-4 inhibitor (preferably linagliptin), optionally in combination with one or more other active agents, for use in treating, preventing, reducing the risk of, slowing progression of, delaying the onset of, and/or protecting against diastolic dysfunction (diastolic heart failure) and/or cardiac fibrosis, and/or diseases associated therewith (e.g. major adverse cardiovascular events), particularly in patients with chronic kidney disease (CKD) such as advancing or severe renal impairment or, particularly, end-stage renal disease (ESRD).

The present invention further relates to a certain DPP-4 inhibitor (preferably linagliptin), optionally in combination with one or more other active agents, for use in preventing, reducing the risk of, slowing progression of, delaying the occurrence of, and/or protecting against cardiovascular morbidity or mortality, such as major adverse cardiovascular events (MACE) (e.g. (non-fatal) myocardial infarction, (non-fatal) stroke, cardiovascular death, or hospitalization for unstable angina pectoris, heart failure or coronary revascularization) in patients with HFpEF and/or CKD (e.g. advancing or severe renal impairment or, particularly, end-stage renal disease (ESRD)).

Further, the invention relates to a method of treating, preventing, reducing the risk of, slowing progression of, delaying the onset of, and/or protecting against heart failure with preserved ejection fraction (HFpEF) or diseases associated therewith (e.g. major adverse cardiovascular events), particularly in a patient (particularly human patient) with chronic kidney disease (CKD) such as advancing or severe renal impairment or end-stage renal disease (ESRD), said method comprising administering an effective amount of a DPP-4 inhibitor, particularly linagliptin, optionally in combination with one or more other therapeutic agents, to the patient in need thereof (particularly human patient).

Further, the present invention relates to the use of a certain DPP-4 inhibitor (preferably linagliptin), optionally in combination with one or more other active agents, for treating, preventing, reducing the risk of, slowing progression of, delaying the onset of, and/or protecting against heart failure with preserved ejection fraction (HFpEF) or diseases associated therewith (e.g. major adverse cardiovascular events), particularly in patients with chronic kidney disease (CKD) such as advancing or severe renal impairment or end-stage renal disease (ESRD).

Further, the present invention relates to the use of a certain DPP-4 inhibitor (preferably linagliptin) for preparing a pharmaceutical composition for treating, preventing, reducing the risk of, slowing progression of, delaying the onset of, and/or protecting against heart failure with preserved ejection fraction (HFpEF) or diseases associated therewith (e.g. major adverse cardiovascular events), particularly in patients with chronic kidney disease (CKD) such as advancing or severe renal impairment or end-stage renal disease (ESRD), optionally in combination with one or more other active agents.

Other aspects of the present invention become apparent to the skilled person from the foregoing and following remarks (including the examples and claims).

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A to 1G show the effects of DPP-4 inhibition on left ventricular (LV) end diastolic pressure volume relationship (EDPVR) in sham and subtotally nephrectomized (SNx) rats:

FIG. 1A shows pressure volume loops for sham+vehicle.

FIG. 1B shows pressure volume loops for sham+linagliptin.

FIG. 1C shows pressure volume loops for sham+sitagliptin.

FIG. 1D shows pressure volume loops for SNx+vehicle.

FIG. 1E shows pressure volume loops for SNx+linagliptin.

FIG. 1F shows pressure volume loops for SNx+sitagliptin.

FIG. 1G shows pressure volume loops for LV EDPVR for each of sham+vehicle, sham+linagliptin, sham+sitagliptin, SNx+vehicle, SNx+linagliptin, and SNx+sitagliptin. (p<0.0001 vs. sham+vehicle, p<0.01 vs. SNx+vehicle).

FIGS. 2A, 2B1, 2B2, 2C1 and 2C2 show the effects of DPP-4 inhibition on cardiac structure in sham and subtotally nephrectomized (SNx) rats:

FIG. 2A shows cardiac collagen I mRNA for each of sham+vehicle, sham+linagliptin, sham+sitagliptin, SNx+vehicle, SNx+linagliptin, and SNx+sitagliptin.

FIG. 2B1 shows representative heart sections immunostained for collagen I from sham+vehicle (B), sham+linagliptin (C), sham+sitagliptin (D), SNx+vehicle (E), SNx+linagliptin (F), and SNx+sitagliptin (G). (p<0.0001 vs. sham+vehicle, p<0.01 vs. SNx+vehicle, p<0.001 vs. SNx+vehicle). Original magnification ×400.

FIG. 2B2 shows the quantitation of cardiac collagen I protein for each of sham+vehicle, sham+linagliptin, sham+sitagliptin, SNx+vehicle, SNx+linagliptin, and SNx+sitagliptin reported in FIG. 2B1. (p<0.01 vs. sham+vehicle, p<0.05 vs. SNx+vehicle).

FIG. 2C1 shows representative H&E-stained heart sections from sham+vehicle (I), sham+linagliptin (J), sham+sitagliptin (K), SNx+vehicle (L), SNx+linagliptin (M), and SNx+sitagliptin (N). Original magnification ×400.

FIG. 2C2 shows the quantitation (myocyte area) of H&E-stained heart sections for each of sham+vehicle, sham+linagliptin, sham+sitagliptin, SNx+vehicle, SNx+linagliptin, and SNx+sitagliptin reported in FIG. 2C1 (p<0.05 vs. sham+vehicle). AU=arbitrary units.

FIG. 3 is a table which includes data for functional, structural and molecular indicators of cardiac dysfunction in subtotally nephrectomized (SNx) rats compared to sham-operated rats.

FIGS. 4a and 4b is a two-part table which includes data for cardiac function in sham and subtotally nephrectomized (SNx) rats treated with vehicle, linagliptin or sitagliptin.

DETAILED DESCRIPTION OF THE INVENTION

Within the scope of the present invention it has been found that certain DPP-4 inhibitors, preferably linagliptin, optionally in combination with one or more other active agents, each as described herein, have properties which make them suitable for the purpose of this invention.

For example, the dipeptidyl peptidase (DPP)-4 inhibitors linagliptin and sitagliptin have been found to prevent the development of cardiac diastolic dysfunction, with cardiac collagen I synthesis also being reduced by DPP-4 inhibition, in a well-established non-diabetic experimental model of CKD and HFpEF, independent from preservation in renal function and/or with renal neutrality, which makes such cardioprotective therapy particularly useful for the patient population with severe renal impairment or ESRD.

It is noteworthy that most DPP-4 inhibitors (e.g. sitagliptin, saxagliptin, alogliptin and vildagliptin) are cleared in the urine and require dose adjustment in the CKD population. The DPP-4 inhibitor linagliptin, however, is unique in being secreted via the bile and does not require adjustment of dose with declining glomerular filtration rate (GFR).

Further, a recent meta-analysis suggested that linagliptin is associated with a reduction in albuminuria in patients with type 2 diabetes.

Additionally, DPP-4 inhibitors can exert anti-fibrotic effects, such as on heart, liver or kidney fibrosis.

In an embodiment, the patient described herein is a diabetic patient (particularly human), such as having diabetes (e.g. type 1 or type 2 diabetes or LADA, particularly type 2 diabetes).

In another embodiment, the patient described herein is a non-diabetic patient (particularly human), such as without diabetes (e.g. without type 1 diabetes, type 2 diabetes or LADA).

In a further embodiment, the patient described herein is a (diabetic or non-diabetic) patient (particularly human patient) with chronic kidney disease (CKD), such as advancing or severe renal impairment or end-stage renal disease (ESRD).

In an embodiment, the patient described herein is a (diabetic or non-diabetic) patient (particularly human patient) with severe renal impairment.

In a particular embodiment, the patient described herein is a (diabetic or non-diabetic) patient (particularly human patient) with end-stage renal disease (ESRD).

The present invention further relates to a pharmaceutical composition or combination comprising or consisting essentially of a certain DPP-4 inhibitor (particularly linagliptin), optionally in combination or alternation with one or more other therapeutic agents, each as described herein, such as e.g. for simultaneous, sequential or separate medical use in therapy or prophylaxis as described herein.

Within this invention it is to be understood that the combinations or combined uses according to this invention may envisage the simultaneous, sequential or separate administration of the therapeutic components.

In this context, “combination” or “combined” within the meaning of this invention may include, without being limited, fixed and non-fixed (e.g. free) forms (including kits, or other administration, application or dosage forms) and uses, such as e.g. the simultaneous, sequential or separate use of the components.

The combined administration or application of this invention may take place by administering the therapeutic components together, such as e.g. by administering them simultaneously in one single or in two separate formulations. Alternatively, the administration may take place by administering the therapeutic components sequentially, such as e.g. successively in two separate formulations.

For the combination therapy of this invention the therapeutic components may be administered separately (which implies that they are formulated separately) or formulated altogether (which implies that they are formulated in the same preparation). Hence, the administration of one element of the combination of the present invention may be prior to, concurrent to, or subsequent to the administration of the other element of the combination.

A DPP-4 inhibitor within the meaning of the present invention includes, without being limited to, any of those DPP-4 inhibitors mentioned hereinabove and hereinbelow, preferably orally active DPP-4 inhibitors. In a further embodiment, a DPP-4 inhibitor within the meaning of the present invention includes orally and/or subcutaneously or transdermally and/or topically active DPP-4 inhibitors.

In a first embodiment (embodiment A), a DPP-4 inhibitor in the context of the present invention is any DPP-4 inhibitor of

wherein R1 denotes ([1,5]naphthyridin-2-yl)methyl, (quinazolin-2-yl)methyl, (quinoxalin-6-yl)methyl, (4-methyl-quinazolin-2-yl)methyl, 2-cyano-benzyl, (3-cyano-quinolin-2-yl)methyl, (3-cyano-pyridin-2-yl)methyl, (4-methyl-pyrimidin-2-yl)methyl, or (4,6-dimethyl-pyrimidin-2-yl)methyl and R2 denotes 3-(R)-amino-piperidin-1-yl, (2-amino-2-methyl-propyl)-methylamino or (2-(S)-amino-propyl)-methylamino,
or its pharmaceutically acceptable salt.

Regarding the first embodiment (embodiment A), preferred DPP-4 inhibitors are any or all of the following compounds and their pharmaceutically acceptable salts:

• 1-[(4-methyl-quinazolin-2-yl)methyl]-3-methyl-7-(2-butyn-1-yl)-8-(3-(R)-amino-piperidin-1-yl)-xanthine (compare WO 2004/018468, example 2(142)):

• 1-[([1,5]naphthyridin-2-yl)methyl]-3-methyl-7-(2-butyn-1-yl)-8-((R)-3-amino-piperidin-1-yl)-xanthine (compare WO 2004/018468, example 2(252)):

• 1-[(Quinazolin-2-yl)methyl]-3-methyl-7-(2-butyn-1-yl)-8-((R)-3-amino-piperidin-1-yl)-xanthine (compare WO 2004/018468, example 2(80)):

• 2-((R)-3-Amino-piperidin-1-yl)-3-(but-2-yinyl)-5-(4-methyl-quinazolin-2-ylmethyl)-3,5-dihydro-imidazo[4,5-d]pyridazin-4-one (compare WO 2004/050658, example 136):

• 1-[(4-Methyl-quinazolin-2-yl)methyl]-3-methyl-7-(2-butyin-1-yl)-8-[(2-amino-2-methyl-propyl)-methylamino]-xanthine (compare WO 2006/029769, example 2(1)):

• 1-[(3-Cyano-quinolin-2-yl)methyl]-3-methyl-7-(2-butyn-1-yl)-8-((R)-3-amino-piperidin-1-yl)-xanthine (compare WO 2005/085246, example 1(30)):

• 1-(2-Cyano-benzyl)-3-methyl-7-(2-butyn-1-yl)-8-((R)-3-amino-piperidin-1-yl)-xanthine (compare WO 2005/085246, example 1(39)):

• 1-[(4-Methyl-quinazolin-2-yl)methyl]-3-methyl-7-(2-butyn-1-yl)-8-[(S)-(2-amino-propyl)-methylamino]-xanthine (compare WO 2006/029769, example 2(4)):

• 1-[(3-Cyano-pyridin-2-yl)methyl]-3-methyl-7-(2-butyn-1-yl)-8-((R)-3-amino-piperidin-1-yl)-xanthine (compare WO 2005/085246, example 1(52)):

• 1-[(4-Methyl-pyrimidin-2-yl)methyl]-3-methyl-7-(2-butyn-1-yl)-8-((R)-3-amino-piperidin-1-yl)-xanthine (compare WO 2005/085246, example 1(81)):

• 1-[(4,6-Dimethyl-pyrimidin-2-yl)methyl]-3-methyl-7-(2-butyn-1-yl)-8-((R)-3-amino-piperidin-1-yl)-xanthine (compare WO 2005/085246, example 1(82)):

• 1-[(Quinoxalin-6-yl)methyl]-3-methyl-7-(2-butyn-1-yl)-8-((R)-3-amino-piperidin-1-yl)-xanthine (compare WO 2005/085246, example 1(83)):

These DPP-4 inhibitors are distinguished from structurally comparable DPP-4 inhibitors, as they combine exceptional potency and a long-lasting effect with favourable pharmacological properties, receptor selectivity and a favourable side-effect profile or bring about unexpected therapeutic advantages or improvements when combined with other pharmaceutical active substances. Their preparation is disclosed in the publications mentioned.

In a second embodiment (embodiment B), a DPP-4 inhibitor in the context of the present invention is a DPP-4 inhibitor selected from the group consisting of sitagliptin, vildagliptin, saxagliptin, alogliptin, gemigliptin, omarigliptin, evogliptin,

• (2S)-1-{[2-(5-Methyl-2-phenyl-oxazol-4-yl)-ethylamino]-acetyl}-pyrrolidine-2-carbonitrile,
• (2S)-1-{[1,1,-Dimethyl-3-(4-pyridin-3-yl-imidazol-1-yl)-propylamino]-acetyl}-pyrrolidine-2-carbonitrile,
• (S)-1-((2S,3S,11bS)-2-Amino-9,10-dimethoxy-1,3,4,6,7,11b-hexahydro-2H-pyrido[2,1-a]isoquinolin-3-yl)-4-fluoromethyl-pyrrolidin-2-one,
• (3,3-Difluoropyrrolidin-1-yl)-((2S,4S)-4-(4-(pyrimidin-2-yl)piperazin-1-yl)pyrrolidin-2-yl)methanone,
• (1((3S,4S)-4-amino-1-(4-(3,3-difluoropyrrolidin-1-yl)-1,3,5-triazin-2-yl)pyrrolidin-3-yl)-5,5-difluoropiperidin-2-one,
• (2S,4S)-1-{2-[(3S,1R)-3-(1H-1,2,4-Triazol-1-ylmethyl)cyclopentylamino]-acetyl}-4-fluoropyrrolidine-2-carbonitrile,
• (R)-2-[6-(3-Amino-piperidin-1-yl)-3-methyl-2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-ylmethyl]-4-fluoro-benzonitrile,
• 5-{(S)-2-[2-((S)-2-Cyano-pyrrolidin-1-yl)-2-oxo-ethylamino]-propyl}-5-(1H-tetrazol-5-yl)-10,11-dihydro-5H-dibenzo[a,d]cycloheptene-2,8-dicarboxylic acid bis-dimethylamide,
• 3-{(2S,4S)-4-[4-(3-Methyl-1-phenyl-1H-pyrazol-5-yl)piperazin-1-yl]pyrrolidin-2-ylcarbonyl}thiazolidine,
• [(2R)-1-{[(3R)-pyrrolidin-3-ylamino]acetyl}pyrrolidin-2-yl]boronic acid,
• (2S,4S)-1-[2-[(4-ethoxycarbonylbicyclo[2.2.2]oct-1-yl)amino]acetyl]-4-fluoropyrrolidine-2-carbonitrile,
• 2-({6-[(3R)-3-amino-3-methylpiperidin-1-yl]-1,3-dimethyl-2,4-dioxo-1,2,3,4-tetrahydro-5H-pyrrolo[3,2-d]pyrimidin-5-yl}methyl)-4-fluorobenzonitrile,
• 6-[(3R)-3-amino-piperidin-1-yl]-5-(2-chloro-5-fluoro-benzyl)-1,3-dimethyl-1,5-dihydro-pyrrolo[3,2-d]pyrimidine-2,4-dione, and
• (S)-2-methylpyrazolo[1,5-a]primidine-6-carboxylic acid {2-[(2-cyanopyrrolidin-1-yl)-2-oxoethylamino]-2-methylpropyl}amide,
  or its pharmaceutically acceptable salt.

A more preferred DPP-4 inhibitor among the abovementioned DPP-4 inhibitors of embodiment A of this invention is 1-[(4-methyl-quinazolin-2-yl)methyl]-3-methyl-7-(2-butyn-1-yl)-8-(3-(R)-amino-piperidin-1-yl)-xanthine, particularly the free base thereof (which is also known as linagliptin or BI 1356).

Preferably the DPP-4 inhibitor of this invention is selected from the group consisting of linagliptin, sitagliptin, vildagliptin, alogliptin, saxagliptin, teneligliptin, anagliptin, gemigliptin and dutogliptin, or a pharmaceutically acceptable salt of one of the herein mentioned DPP-4 inhibitors, or a prodrug thereof.

A particularly preferred DPP-4 inhibitor to be emphasized within the present invention is linagliptin. The term “linagliptin” as employed herein refers to linagliptin or a pharmaceutically acceptable salt thereof, including hydrates and solvates thereof, and crystalline forms thereof, preferably linagliptin refers to 1-[(4-methyl-quinazolin-2-yl)methyl]-3-methyl-7-(2-butyn-1-yl)-8-(3-(R)-amino-piperidin-1-yl)-xanthine. Crystalline forms are described in WO 2007/128721. Methods for the manufacture of linagliptin are described in the patent applications WO 2004/018468 and WO 2006/048427 for example. Linagliptin is distinguished from structurally comparable DPP-4 inhibitors, as it combines exceptional potency and a long-lasting effect with favourable pharmacological properties, receptor selectivity and a favourable side-effect profile or bring about unexpected therapeutic advantages or improvements in therapy.

With respect to embodiment A, the methods of synthesis for the DPP-4 inhibitors according to embodiment A of this invention are known to the skilled person. Advantageously, the DPP-4 inhibitors according to embodiment A of this invention can be prepared using synthetic methods as described in the literature. Thus, for example, purine derivatives of formula (I) can be obtained as described in WO 2002/068420, WO 2004/018468, WO 2005/085246, WO 2006/029769 or WO 2006/048427, the disclosures of which are incorporated herein. Purine derivatives of formula (II) can be obtained as described, for example, in WO 2004/050658 or WO 2005/110999, the disclosures of which are incorporated herein. Purine derivatives of formula (III) and (IV) can be obtained as described, for example, in WO 2006/068163, WO 2007/071738 or WO 2008/017670, the disclosures of which are incorporated herein. The preparation of those DPP-4 inhibitors, which are specifically mentioned hereinabove, is disclosed in the publications mentioned in connection therewith. Polymorphous crystal modifications and formulations of particular DPP-4 inhibitors are disclosed in WO 2007/128721 and WO 2007/128724, respectively, the disclosures of which are incorporated herein in their entireties. Formulations of particular DPP-4 inhibitors with metformin or other combination partners are described in WO 2009/121945, the disclosure of which is incorporated herein in its entirety.

With respect to embodiment B, the methods of synthesis for the DPP-4 inhibitors of embodiment B are described in the scientific literature and/or in published patent documents, particularly in those cited herein.

In an embodiment, the DPP-4 inhibitor according to the invention is preferably administered orally.

Suitable doses and dosage forms of the DPP-4 inhibitors may be determined by a person skilled in the art and may include those described herein or in the relevant references.

For pharmaceutical application in warm-blooded vertebrates, particularly humans, the compounds of this invention are usually used in dosages from 0.001 to 100 mg/kg body weight, preferably at 0.01-15 mg/kg or 0.1-15 mg/kg, in each case 1 to 4 times a day. For this purpose, the compounds, optionally combined with other active substances, may be incorporated together with one or more inert conventional carriers and/or diluents, e.g. with corn starch, lactose, glucose, microcrystalline cellulose, magnesium stearate, polyvinylpyrrolidone, citric acid, tartaric acid, water, water/ethanol, water/glycerol, water/sorbitol, water/polyethylene glycol, propylene glycol, cetylstearyl alcohol, carboxymethylcellulose or fatty substances such as hard fat or suitable mixtures thereof into conventional galenic preparations such as plain or coated tablets, capsules, powders, suspensions or suppositories.

The pharmaceutical compositions according to this invention comprising the DPP-4 inhibitors as defined herein are thus prepared by the skilled person using pharmaceutically acceptable formulation excipients as described in the art and appropriate for the desired route of administration. Examples of such excipients include, without being restricted to diluents, binders, carriers, fillers, lubricants, flow promoters, crystallisation retardants, disintegrants, solubilizers, colorants, pH regulators, surfactants and emulsifiers.

Oral formulations or dosage forms of the DPP-4 inhibitor of this invention may be prepared according to known techniques.

A pharmaceutical composition or dosage form (e.g. oral tablet) of a DPP-4 inhibitor according to embodiment A of the invention may typically contain as excipients (in addition to an active ingredient), for example: one or more diluents, a binder, a disintegrant, and a lubricant, preferably each as disclosed herein-below. In an embodiment, the disintegrant may be optional.

Examples of suitable diluents for compounds according to embodiment A include cellulose powder, calcium hydrogen phosphate, erythritol, low substituted hydroxypropyl cellulose, mannitol, pregelatinized starch or xylitol.

Examples of suitable lubricants for compounds according to embodiment A include talc, polyethyleneglycol, calcium behenate, calcium stearate, hydrogenated castor oil or magnesium stearate.

Examples of suitable binders for compounds according to embodiment A include copovidone (copolymerisates of vinylpyrrolidon with other vinylderivates), hydroxypropyl methylcellulose (HPMC), hydroxypropylcellulose (HPC), polyvinylpyrrolidon (povidone), pregelatinized starch, or low-substituted hydroxypropylcellulose (L-HPC).

Examples of suitable disintegrants for compounds according to embodiment A include corn starch or crospovidone.

Suitable methods of preparing (oral) preparations or dosage forms of the DPP-4 inhibitors according to embodiment A of the invention are:

direct tabletting of the active substance in powder mixtures with suitable tabletting excipients;

granulation with suitable excipients and subsequent mixing with suitable excipients and subsequent tabletting as well as film coating; or

packing of powder mixtures or granules into capsules.

Suitable granulation methods are:

• wet granulation in the intensive mixer followed by fluidised bed drying;
• one-pot granulation;
• fluidised bed granulation; or
• dry granulation (e.g. by roller compaction) with suitable excipients and subsequent tabletting or packing into capsules.

An exemplary composition for oral use (e.g. tablet core) of a DPP-4 inhibitor according to embodiment A of the invention comprises the first diluent mannitol, pregelatinized starch as a second diluent with additional binder properties, the binder copovidone, the disintegrant corn starch, and magnesium stearate as lubricant; wherein copovidone and/or corn starch may be optional.

A tablet of a DPP-4 inhibitor according to embodiment A of the invention may be film coated, preferably the film coat comprises hydroxypropylmethylcellulose (HPMC), polyethylene glycol (PEG), talc, titanium dioxide and iron oxide (e.g. red and/or yellow).

For details on dosage forms, formulations and administration of DPP-4 inhibitors of this invention, reference is made to scientific literature and/or published patent documents, particularly to those cited herein.

With respect to the first embodiment (embodiment A), the dosage typically required of the DPP-4 inhibitors mentioned herein in embodiment A when administered intravenously is 0.1 mg to 10 mg, preferably 0.25 mg to 5 mg, and when administered orally is 0.5 mg to 100 mg, preferably 2.5 mg to 50 mg or 0.5 mg to 10 mg, more preferably 2.5 mg to 10 mg or 1 mg to 5 mg, in each case 1 to 4 times a day. Thus, e.g. the dosage of 1-[(4-methyl-quinazolin-2-yl)methyl]-3-methyl-7-(2-butyn-1-yl)-8-(3-(R)-amino-piperidin-1-yl)-xanthine when administered orally is 0.5 mg to 10 mg per patient per day, preferably 2.5 mg to 10 mg or 1 mg to 5 mg per patient per day.

A dosage form prepared with a pharmaceutical composition comprising a DPP-4 inhibitor mentioned herein in embodiment A contain the active ingredient in a dosage range of 0.1-100 mg. Thus, e.g. particular oral dosage strengths of 1-[(4-methyl-quinazolin-2-yl)methyl]-3-methyl-7-(2-butyn-1-yl)-8-(3-(R)-amino-piperidin-1-yl)-xanthine are 0.5 mg, 1 mg, 2.5 mg, 5 mg and 10 mg.

A special embodiment of the DPP-4 inhibitors of this invention refers to those orally administered DPP-4 inhibitors which are therapeutically efficacious at low dose levels, e.g. at oral dose levels <100 mg or <70 mg per patient per day, preferably <50 mg, more preferably <30 mg or <20 mg, even more preferably from 1 mg to 10 mg, particularly from 1 mg to 5 mg (more particularly 5 mg), per patient per day (if required, divided into 1 to 4 single doses, particularly 1 or 2 single doses, which may be of the same size, preferentially, administered orally once- or twice daily (more preferentially once-daily), advantageously, administered at any time of day, with or without food. Thus, for example, the daily oral amount 5 mg BI 1356 can be given in an once daily dosing regimen (i.e. 5 mg BI 1356 once daily) or in a twice daily dosing regimen (i.e. 2.5 mg BI 1356 twice daily), at any time of day, with or without food.

The dosage of the active components in the combinations or compositions in accordance with the present invention may be varied, although the amount of the active ingredients shall be such that a suitable dosage form is obtained. Hence, the selected dosage and the selected dosage form shall depend on the desired therapeutic effect, the route of administration and the duration of the treatment. Dosage ranges for the combination may be from the maximal tolerated dose for the single agent to lower doses.

The present invention further provides a certain DPP-4 inhibitor as defined herein (preferably linagliptin, optionally in combination with one or more other active agents) for use in for treating and/or preventing metabolic diseases, particularly diabetes, especially type 2 diabetes mellitus, and/or conditions related thereto (e.g. diabetic complications), in a patient (particularly human patient) having or being at risk of HFpEF and/or chronic kidney disease (CKD) such as advancing or severe renal impairment or end-stage renal disease (ESRD).

Examples of metabolic disorders or diseases amenable by the therapy of this invention may include, without being limited to, type 1 diabetes, type 2 diabetes, impaired glucose tolerance (IGT), impaired fasting blood glucose (IFG), hyperglycemia, postprandial hyperglycemia, postabsorptive hyperglycemia, latent autoimmune diabetes in adults (LADA), overweight, obesity, dyslipidemia, hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, hyperNEFA-emia, fasting or postprandial hyperlipidemia such as postprandial lipemia (e.g. postprandial hypertriglyceridemia), hypertension, atherosclerosis, endothelial dysfunction, osteoporosis, chronic systemic inflammation, non alcoholic fatty liver disease (NAFLD), retinopathy, neuropathy, nephropathy, polycystic ovarian syndrome, and/or metabolic syndrome.

The present invention further relates to a certain DPP-4 inhibitor (preferably linagliptin, optionally in combination with one or more other active agents) for use in at least one of the following methods:

• • preventing, slowing the progression of, delaying the onset of or treating a metabolic disorder or disease, such as e.g. type 1 diabetes mellitus, type 2 diabetes mellitus, impaired glucose tolerance (IGT), impaired fasting blood glucose (IFG), hyperglycemia, postprandial hyperglycemia, postabsorptive hyperglycemia, latent autoimmune diabetes in adults (LADA), overweight, obesity, dyslipidemia, hyperlipidemia, hypercholesterolemia, hypertriglyceridemia, hyperNEFA-emia, postprandial lipemia (e.g. postprandial hypertriglyceridemia), hypertension, atherosclerosis, endothelial dysfunction, osteoporosis, chronic systemic inflammation, non alcoholic fatty liver disease (NAFLD), retinopathy, neuropathy, nephropathy, polycystic ovarian syndrome, and/or metabolic syndrome;
  • improving and/or maintaining glycemic control and/or for reducing of fasting plasma glucose, of postprandial plasma glucose, of postabsorptive plasma glucose and/or of glycosylated hemoglobin HbA1c, or preventing, reducing the risk of, slowing the progression of, delaying the onset of or treating worsening or deterioration of glycemic control, need for insulin therapy or elevated HbA1c despite treatment;
  • preventing, slowing, delaying the onset of or reversing progression from pre-diabetes, impaired glucose tolerance (IGT), impaired fasting blood glucose (IFG), insulin resistance and/or from metabolic syndrome to type 2 diabetes mellitus;
  • preventing, reducing the risk of, slowing the progression of, delaying the onset of or treating of complications of diabetes mellitus such as micro- and macrovascular diseases, such as nephropathy, micro- or macroalbuminuria, proteinuria, retinopathy, cataracts, neuropathy, learning or memory impairment, neurodegenerative or cognitive disorders, cardio- or cerebrovascular diseases, tissue ischaemia, diabetic foot or ulcus, atherosclerosis, hypertension, endothelial dysfunction, myocardial infarction, acute coronary syndrome, unstable angina pectoris, stable angina pectoris, peripheral arterial occlusive disease, cardiomyopathy, heart failure, heart rhythm disorders, vascular restenosis, and/or stroke;
  • reducing body weight and/or body fat and/or liver fat and/or intra-myocellular fat or preventing an increase in body weight and/or body fat and/or liver fat and/or intra-myocellular fat or facilitating a reduction in body weight and/or body fat and/or liver fat and/or intra-myocellular fat;
  • preventing, slowing, delaying the onset of or treating the degeneration of pancreatic beta cells and/or the decline of the functionality of pancreatic beta cells and/or for improving, preserving and/or restoring the functionality of pancreatic beta cells and/or stimulating and/or restoring or protecting the functionality of pancreatic insulin secretion;
  • preventing, slowing, delaying the onset of or treating non alcoholic fatty liver disease (NAFLD) including hepatic steatosis, non-alcoholic steatohepatitis (NASH) and/or liver fibrosis (such as e.g. preventing, slowing the progression, delaying the onset of, attenuating, treating or reversing hepatic steatosis, (hepatic) inflammation and/or an abnormal accumulation of liver fat);
  • preventing, slowing the progression of, delaying the onset of or treating type 2 diabetes with failure to conventional antidiabetic mono- or combination therapy;
  • achieving a reduction in the dose of conventional antidiabetic medication (e.g. of a sulphonylurea or an insulin) required for adequate therapeutic effect;
  • reducing the risk for adverse effects associated with conventional antidiabetic medication (e.g. hypoglycemia or weight gain, such as associated with e.g. insulin or sulphonylurea medication); and/or
  • maintaining and/or improving the insulin sensitivity and/or for treating or preventing hyperinsulinemia and/or insulin resistance;
    in a patient in need thereof (such as e.g. a patient as described herein), particularly
    in a patient (particularly human patient) with HFpEF and/or chronic kidney disease (CKD) such as advancing or severe renal impairment or end-stage renal disease (ESRD).

As different metabolic or functional disorders often occur simultaneously, it is quite often indicated to combine a number of different active principles with one another. Thus, depending on the functional disorders diagnosed, improved treatment outcomes may be obtained if a DPP-4 inhibitor is combined with one or more active substances customary for the respective disorders, such as e.g. one or more active substances selected from among the other antidiabetic substances, especially active substances that lower the blood sugar level or the lipid level in the blood, raise the HDL level in the blood, lower blood pressure or are indicated in the treatment of atherosclerosis or obesity.

The DPP-4 inhibitors mentioned above—besides their use in mono-therapy—may also be used in conjunction with other active substances, by means of which improved treatment results can be obtained. Such a combined treatment may be given as a free combination of the substances or in the form of a fixed combination, for example in a tablet or capsule. Pharmaceutical formulations of the combination partner needed for this may either be obtained commercially as pharmaceutical compositions or may be formulated by the skilled man using conventional methods. The active substances which may be obtained commercially as pharmaceutical compositions are described in numerous places in the prior art, for example in the list of drugs that appears annually, the “Rote Liste®” of the federal association of the pharmaceutical industry, or in the annually updated compilation of manufacturers' information on prescription drugs known as the “Physicians' Desk Reference”.

Examples of antidiabetic combination partners are metformin; sulphonylureas such as glibenclamide, tolbutamide, glimepiride, glipizide, gliquidon, glibornuride and gliclazide; nateglinide; repaglinide; mitiglinide; thiazolidinediones such as rosiglitazone and pioglitazone; PPAR gamma modulators such as metaglidases; PPAR-gamma agonists such as e.g. rivoglitazone, mitoglitazone, INT-131 and balaglitazone; PPAR-gamma antagonists; PPAR-gamma/alpha modulators such as tesaglitazar, muraglitazar, aleglitazar, indeglitazar, saroglitazar and KRP297; PPAR-gamma/alpha/delta modulators such as e.g. lobeglitazone; AMPK-activators such as AICAR; acetyl-CoA carboxylase (ACC1 and ACC2) inhibitors; diacylglycerol-acetyltransferase (DGAT) inhibitors; pancreatic beta cell GCRP agonists such as GPR119 agonists (SMT3-receptor-agonists); 11β-HSD-inhibitors; FGF19 agonists or analogues; alpha-glucosidase blockers such as acarbose, voglibose and miglitol; alpha2-antagonists; insulin and insulin analogues such as human insulin, insulin lispro, insulin glusilin, r-DNA-insulinaspart, NPH insulin, insulin detemir, insulin degludec, insulin tregopil, insulin zinc suspension and insulin glargin; Gastric inhibitory Peptide (GIP); amylin and amylin analogues (e.g. pramlintide or davalintide); GLP-1 and GLP-1 analogues such as Exendin-4, e.g. exenatide, exenatide LAR, liraglutide, taspoglutide, lixisenatide (AVE-0010), LY-2428757 (a PEGylated version of GLP-1), dulaglutide (LY-2189265), semaglutide or albiglutide; SGLT2-inhibitors such as e.g. dapagliflozin, sergliflozin (KGT-1251), atigliflozin, canagliflozin, ipragliflozin, luseogliflozin or tofogliflozin; inhibitors of protein tyrosine-phosphatase (e.g. trodusquemine); inhibitors of glucose-6-phosphatase; fructose-1,6-bisphosphatase modulators; glycogen phosphorylase modulators; glucagon receptor antagonists; phosphoenolpyruvatecarboxykinase (PEPCK) inhibitors; pyruvate dehydrogenasekinase (PDK) inhibitors; inhibitors of tyrosine-kinases (50 mg to 600 mg) such as PDGF-receptor-kinase (cf. EP-A-564409, WO 98/35958, U.S. Pat. No. 5,093,330, WO 2004/005281, and WO 2006/041976) or of serine/threonine kinases; glucokinase/regulatory protein modulators incl. glucokinase activators; glycogen synthase kinase inhibitors; inhibitors of the SH2-domain-containing inositol 5-phosphatase type 2 (SHIP2); IKK inhibitors such as high-dose salicylate; JNK1 inhibitors; protein kinase C-theta inhibitors; beta 3 agonists such as ritobegron, YM 178, solabegron, talibegron, N-5984, GRC-1087, rafabegron, FMP825; aldosereductase inhibitors such as AS 3201, zenarestat, fidarestat, epalrestat, ranirestat, NZ-314, CP-744809, and CT-112; SGLT-1 or SGLT-2 inhibitors; KV 1.3 channel inhibitors; GPR40 modulators such as e.g. [(35)-6-({2′,6′-dimethyl-4′-[3-(methylsulfonyl)propoxy]biphenyl-3-yl}methoxy)-2,3-dihydro-1-benzofuran-3-yl]acetic acid; SCD-1 inhibitors; CCR-2 antagonists; dopamine receptor agonists (bromocriptine mesylate [Cycloset]); 4-(3-(2,6-dimethylbenzyloxyl)phenyl)-4-oxobutanoic acid; sirtuin stimulants; and other DPP IV inhibitors.

Metformin is usually given in doses varying from about 500 mg to 2000 mg up to 2500 mg per day using various dosing regimens from about 100 mg to 500 mg or 200 mg to 850 mg (1-3 times a day), or about 300 mg to 1000 mg once or twice a day, or delayed-release metformin in doses of about 100 mg to 1000 mg or preferably 500 mg to 1000 mg once or twice a day or about 500 mg to 2000 mg once a day. Particular dosage strengths may be 250, 500, 625, 750, 850 and 1000 mg of metformin hydrochloride.

A dosage of pioglitazone is usually of about 1-10 mg, 15 mg, 30 mg, or 45 mg once a day.

Rosiglitazone is usually given in doses from 4 to 8 mg once (or divided twice) a day (typical dosage strengths are 2, 4 and 8 mg).

Glibenclamide (glyburide) is usually given in doses from 2.5-5 to 20 mg once (or divided twice) a day (typical dosage strengths are 1.25, 2.5 and 5 mg), or micronized glibenclamide in doses from 0.75-3 to 12 mg once (or divided twice) a day (typical dosage strengths are 1.5, 3, 4.5 and 6 mg).

Glipizide is usually given in doses from 2.5 to 10-20 mg once (or up to 40 mg divided twice) a day (typical dosage strengths are 5 and 10 mg), or extended-release glibenclamide in doses from 5 to 10 mg (up to 20 mg) once a day (typical dosage strengths are 2.5, 5 and 10 mg). Glimepiride is usually given in doses from 1-2 to 4 mg (up to 8 mg) once a day (typical dosage strengths are 1, 2 and 4 mg).

• A dual combination of glibenclamide/metformin is usually given in doses from 1.25/250 once daily to 10/1000 mg twice daily. (typical dosage strengths are 1.25/250, 2.5/500 and 5/500 mg).
• A dual combination of glipizide/metformin is usually given in doses from 2.5/250 to 10/1000 mg twice daily (typical dosage strengths are 2.5/250, 2.5/500 and 5/500 mg).

A dual combination of glimepiride/metformin is usually given in doses from 1/250 to 4/1000 mg twice daily.

• A dual combination of rosiglitazone/glimepiride is usually given in doses from 4/1 once or twice daily to 4/2 mg twice daily (typical dosage strengths are 4/1, 4/2, 4/4, 8/2 and 8/4 mg).
• A dual combination of pioglitazone/glimepiride is usually given in doses from 30/2 to 30/4 mg once daily (typical dosage strengths are 30/4 and 45/4 mg).
• A dual combination of rosiglitazone/metformin is usually given in doses from 1/500 to 4/1000 mg twice daily (typical dosage strengths are 1/500, 2/500, 4/500, 2/1000 and 4/1000 mg).
• A dual combination of pioglitazone/metformin is usually given in doses from 15/500 once or twice daily to 15/850 mg thrice daily (typical dosage strengths are 15/500 and 15/850 mg).

The non-sulphonylurea insulin secretagogue nateglinide is usually given in doses from 60 to 120 mg with meals (up to 360 mg/day, typical dosage strengths are 60 and 120 mg); repaglinide is usually given in doses from 0.5 to 4 mg with meals (up to 16 mg/day, typical dosage strengths are 0.5, 1 and 2 mg). A dual combination of repaglinide/metformin is available in dosage strengths of 1/500 and 2/850 mg.

Acarbose is usually given in doses from 25 to 100 mg with meals. Miglitol is usually given in doses from 25 to 100 mg with meals.

Examples of combination partners that lower the lipid level in the blood are HMG-CoA-reductase inhibitors such as simvastatin, atorvastatin, lovastatin, fluvastatin, pravastatin, pitavastatin and rosuvastatin; fibrates such as bezafibrate, fenofibrate, clofibrate, gemfibrozil, etofibrate and etofyllinclofibrate; nicotinic acid and the derivatives thereof such as acipimox; PPAR-alpha agonists; PPAR-delta agonists such as e.g. {4-[(R)-2-ethoxy-3-(4-trifluoromethyl-phenoxy)-propylsulfanyl]-2-methyl-phenoxy}-acetic acid; PPAR-alpha/delta agonists; inhibitors of acyl-coenzyme A:cholesterolacyltransferase (ACAT; EC 2.3.1.26) such as avasimibe; cholesterol resorption inhibitors such as ezetimib; substances that bind to bile acid, such as cholestyramine, colestipol and colesevelam; inhibitors of bile acid transport; HDL modulating active substances such as D4F, reverse D4F, LXR modulating active substances and FXR modulating active substances; CETP inhibitors such as torcetrapib, JTT-705 (dalcetrapib) or compound 12 from WO 2007/005572 (anacetrapib); LDL receptor modulators; MTP inhibitors (e.g. lomitapide); and ApoB100 antisense RNA.

A dosage of atorvastatin is usually from 1 mg to 40 mg or 10 mg to 80 mg once a day.

Examples of combination partners that lower blood pressure are beta-blockers such as atenolol, bisoprolol, celiprolol, metoprolol and carvedilol; diuretics such as hydrochlorothiazide, chlortalidon, xipamide, furosemide, piretanide, torasemide, spironolactone, eplerenone, amiloride and triamterene; calcium channel blockers such as amlodipine, nifedipine, nitrendipine, nisoldipine, nicardipine, felodipine, lacidipine, lercanipidine, manidipine, isradipine, nilvadipine, verapamil, gallopamil and diltiazem; ACE inhibitors such as ramipril, lisinopril, cilazapril, quinapril, captopril, enalapril, benazepril, perindopril, fosinopril and trandolapril; as well as angiotensin II receptor blockers (ARBs) such as telmisartan, candesartan, valsartan, losartan, irbesartan, olmesartan, azilsartan and eprosartan.

A dosage of telmisartan is usually from 20 mg to 320 mg or 40 mg to 160 mg per day.

Examples of combination partners which increase the HDL level in the blood are Cholesteryl Ester Transfer Protein (CETP) inhibitors; inhibitors of endothelial lipase; regulators of ABC1; LXRalpha antagonists; LXRbeta agonists; PPAR-delta agonists; LXRalpha/beta regulators, and substances that increase the expression and/or plasma concentration of apolipoprotein A-I.

Examples of combination partners for the treatment of obesity are sibutramine; tetrahydrolipstatin (orlistat); alizyme (cetilistat); dexfenfluramine; axokine; cannabinoid receptor 1 antagonists such as the CB1 antagonist rimonobant; MCH-1 receptor antagonists; MC4 receptor agonists; NPY5 as well as NPY2 antagonists (e.g. velneperit); beta3-AR agonists such as SB-418790 and AD-9677; 5HT2c receptor agonists such as APD 356 (lorcaserin); myostatin inhibitors; Acrp30 and adiponectin; steroyl CoA desaturase (SCD1) inhibitors; fatty acid synthase (FAS) inhibitors; CCK receptor agonists; Ghrelin receptor modulators; Pyy 3-36; orexin receptor antagonists; and tesofensine; as well as the dual combinations bupropion/naltrexone, bupropion/zonisamide, topiramate/phentermine and pramlintide/metreleptin.

Examples of combination partners for the treatment of atherosclerosis are phospholipase A2 inhibitors; inhibitors of tyrosine-kinases (50 mg to 600 mg) such as PDGF-receptor-kinase (cf. EP-A-564409, WO 98/35958, U.S. Pat. No. 5,093,330, WO 2004/005281, and WO 2006/041976); oxLDL antibodies and oxLDL vaccines; apoA-1 Milano; ASA; and VCAM-1 inhibitors.

Further, the certain DPP-4 inhibitor of this invention may be used in combination with a substrate of DPP-4 (particularly with an anti-inflammatory substrate of DPP-4), which may be other than GLP-1, for the purposes according to the present invention, such substrates of DPP-4 include, for example—without being limited to, one or more of the following:

• Incretins:
• Glucagon-like peptide (GLP)-1
• Glucose-dependent insulinotropic peptide (GIP)
• Neuroactive:
• Substance P
• Neuropeptide Y (NPY)
• Peptide YY
• Energy homeostasis:
• GLP-2
• Prolactin
• Pituitary adenylate cyclase activating peptide (PACAP)
• Other hormones:
• PACAP 27
• Human chorionic gonadotrophin alpha chain
• Growth hormone releasing factor (GHRF)
• Luteinizing hormone alpha chain
• Insulin-like growth factor (IGF-1)
• CCL8/eotaxin
• CCL22/macrophage-derived chemokine
• CXCL9/interferon-gamma-induced monokine
• Chemokines:
• CXCL10/interferon-gamma-induced protein-10
• CXCL11/interferon-inducible T cell a chemoattractant
• CCL3L1/macrophage inflammatory protein lalpha isoform
• LD78beta
• CXCL12/stromal-derived factor 1 alpha and beta
• Other:
• Enkephalins, gastrin-releasing peptide, vasostatin-1,
• peptide histidine methionine, thyrotropin alpha

Further or in addition, the certain DPP-4 inhibitor of this invention may be used in combination with one or more active substances which may be typical options in the treatment of HFpEF, such as selected from diuretics, ACE inhibitors, ARBs, beta-blockers, calcium-channel blockers, digoxin and/or statins, and/or agents that modulate cGMP signaling pathway, PDE V inhibitors, neprilysin inhibitors, nitric oxide donors, sGC stimulators and/or aldosterone receptor agonists.

Further or in addition, the certain DPP-4 inhibitor of this invention may be used in combination with one or more active substances which are indicated in the treatment or prevention of cardiovascular diseases or events (e.g. major cardiovascular events).

Moreover, optionally in addition, the certain DPP-4 inhibitor of this invention may be used in combination with one or more antiplatelet agents, such as e.g. (low-dose) aspirin (acetylsalicylic acid), a selective COX-2 or nonselective COX-1/COX-2 inhibitor, or a ADP receptor inhibitor, such as a thienopyridine (e.g. clopidogrel or prasugrel), elinogrel or ticagrelor, or a thrombin receptor antagonist such as vorapaxar.

Yet moreover, optionally in addition, the certain DPP-4 inhibitor of this invention may be used in combination with one or more anticoagulant agents, such as e.g. a heparin, a coumarin (such as warfarin or phenprocoumon), a direct thrombin inhibitor (such as e.g. dabigatran), a pentasaccharide inhibitor of Factor Xa (e.g. fondaparinux) or a direct Faktor Xa inhibitor (such as e.g. rivaroxaban or apixaban or edoxaban or otamixaban).

Still yet moreover, optionally in addition, the certain DPP-4 inhibitor of this invention may be used in combination with one or more agents for the treatment of heart failure.

All patent applications cited herein are hereby incorporated by reference in their entireties.

The present invention is not to be limited in scope by the specific embodiments described herein. Various modifications of the invention in addition to those described herein may become apparent to those skilled in the art from the present disclosure. Such modifications are intended to fall within the scope of the present invention.

Further embodiments, features and advantages of the present invention may become apparent from the following examples. The following examples serve to illustrate, by way of example, the principles of the invention without restricting it.

EXAMPLES

Animals

Female Fischer 344 rats (F344, Charles River, Montreal, Quebec) aged 12 weeks underwent sham or subtotal nephrectomy surgery as previously described.

Briefly, animals were anesthetized with 2.5% inhaled isoflurane, the right kidney was removed via subcapsular nephrectomy and infarction of approximately two thirds of the left kidney was achieved via selective ligation of two out of the three or four branches of the left renal artery. Sham surgery consisted of laparotomy and manipulation of both kidneys before wound closure. Rats were followed for eight weeks after surgery. Ten days post-surgery, rats were randomized to receive either vehicle (0.5% aqueous hydroxyethylcellulose, Sigma), 1.5 mg/kg/day linagliptin (in 0.5% aqueous hydroxyethylcellulose, Boehringer Ingelheim) or 20 mg/kg/day sitagliptin (in 0.25% carboxymethylcellulose, Sequoia Research Products Ltd., Pangbourne, UK) by daily oral gavage for seven weeks. The linagliptin dosage of 1.5 mg/kg is approximately equivalent to 3.2 μmol/kg, midway between two previous doses of the DPP-4 inhibitor employed in an earlier short-term study in SNx rats. All rats were housed in a stable environment and allowed free access to Reverse Osmosis water and standard rat chow.

Metabolic Parameters

Blood glucose was measured with Accu-check Advantage (Roche, Mississauga, Ontario). HbA1c was measured using A1cNow+ (Bayer, Sunnyvale, Calif.). Active GLP-1 was determined using active GLP-1 (v2) kit (Meso Scale Discovery, Rockville, Md.).

Cardiac Function

Transthoracic echocardiography was performed in anesthetized rats (1% isoflurane) using a Sonos 5500 echocardiograph (Philips Healthcare, Andover, Mass.) with a highfrequency (5-12 MHz broad-bandwidth) phased array transducer (S12, Philips). Parasternal long axis and short axis views at the mid-level of the papillary muscle and linear dimensions were analyzed offline by a single investigator masked to the treatment groups. Fractional shortening (FS %) was calculated according to the formula: FS %=(LVIDd−LVIDs)/LVIDd×100, where LVIDd and LVIDs are end-diastolic diameter and end-systolic diameter respectively, as previously described. Left ventricular mass was derived as previously reported. Three consecutive cardiac cycles were averaged for all analyses.

Cardiac catheterization was performed as previously published. Briefly, rats were anesthetized (2% isoflurane), placed on a warming pad (37oC), intubated and ventilated using positive pressure. The right internal carotid artery was ligated cranially and a 2F miniaturized combined conductance catheter-micromanometer (Model SPR-838, Millar Instruments, Houston, Tex.) was inserted into the carotid artery and advanced into the left ventricle until stable pressure volume (PV) loops were obtained. The abdomen was opened and elastic bands were placed around the inferior vena cava and portal vein. All PV loops were obtained with the ventilator turned off for 5-10 seconds and the animal apneic. Data were acquired under steady state conditions and during preload reduction with parallel conductance values obtained by injection of approximately 200 μl 10% NaCl into the right atrium. Calibration from Relative Volume Units (RVU) conductance signal to absolute volumes (in μl) was undertaken using a previously validated method of comparison to known volumes in Perspex wells. A range of functional parameters were then calculated using the pressure conductance data (Millar analysis software PVAN 3.4).

Renal Function

For estimation of urine protein excretion, rats were individually housed in metabolic cages for 24 h with free access to tap water and standard diet. Urine protein was measured with the benzethonium chloride method. Urinary excretion of markers of renal injury was determined using Rat KidneyMAP v1.0 (Myriad RBM, Austin, Tex.) and expressed as the ratio to urine creatinine determined by autoanalyzer. Glomerular filtration rate (GFR) was determined by FITC-inulin clearance, as previously described.

Tissue Collection

Rats were anesthetized (2.5% isoflurane) before cervical dislocation. Cardiac tissue was either fixed in 10% NBF or flash frozen in liquid nitrogen before storage at −80° C. for future molecular biological analysis. The left renal artery was clamped and the kidney was removed, weighed, sliced transversely and immersed in 10% NBF for 24 h. Formalin-fixed tissues were routinely processed, embedded in paraffin and sectioned.

Plasma DPP-4 Activity

Plasma DPP-4 activity was determined in rats (n=3/group) treated with either vehicle, linagliptin or sitagliptin for four days, 1 h and 24 h post-gavage, as previously described. Briefly, 20 μl EDTA plasma was diluted with 30 μl DPP-4 assay buffer (100 mM Tris, 100 mM NaCl, pH 7.8) and 50 μl substrate (100 μM H-Ala-Pro-7-amido-4-trifluromethylcoumarin [AlaPro-AFC], final concentration 100 μM, Bachem, Torrance, Calif.). After 10 min at room temperature, fluorescence was determined using a SpectraMax M5e (Molecular Devices, Downingtown, Pa.), excitation 405 nm, emission 535 nm.

Cardiac Structure

Cardiac Gene Expression

RNA was isolated from homogenized rat heart tissue using TRIzol reagent (Life Technologies, Grand Island, N.Y.). Total RNA (4 μg) was treated with RQ1 DNase (1U/μl) (Promega, Madison, Wis.). DNase treated RNA (4 μg) was reverse-transcribed in a final volume of 25 μl using 0.5 μl AMV-RT (Roche Diagnostics, Laval, Quebec) in the manufacturer's buffer containing 1 mmol/L dNTPs, 0.5 μl RNase inhibitor (Roche) and 2 μg random hexamers (GE Healthcare Life Sciences, Baie d′Urfe, Quebec). SYBR green based real time PCR was performed on an ABI Prism 7900HT Fast PCR System (Applied Biosystems, Foster City, Calif.) using the following primer sequences: (SEQ ID Nos. 1-10) collagen I forward TGCCGATGTCGCTATCCA, reverse TCTTGCAGTGATAGGTGATGTTCTG; α-myosin heavy chain (MHC) forward TGTGAAAAGATTAACCGGAGTTTAAG, reverse TCTGACTTGCGGAGGTATCG; β-MHC forward GTGCCAAGGGCCTGAATGAG, reverse GCAAAGGCTCCAGGTCTGA; atrial natriuretic peptide (ANP) forward ATGGGCTCCTTCTCCATCAC, reverse TCTTCGGTACCGGAAGCT; RPL13a forward GATGAACACCAACCCGTCTC, reverse CACCATCCGCTTTTTCTTGT. Brain natriuretic peptide (BNP) primers were from Qiagen (Germantown, Md.). Data analysis was performed using Applied Biosystems Comparative CT method.

Immunohistochemistry for Cardiac Collagen I

Immunohistochemistry for fibrillar collagen type I (Southern Biotech, Birmingham, Ala.) was performed as previously described with the antibody used at a concentration of 1:400. Stained heart sections were scanned with the Aperio ScanScope system (Aperio, Vista, Calif.) and analysed with ImageScope (Aperio). Interstitial collagen I was determined as the proportional area of positive immunostaining in 10 randomly selected fields (×100 magnification).

Myocyte Hypertrophy

Cardiac myocyte hypertrophy was determined on haematoxylin and eosin (H&E) stained sections, as previously reported. Myocyte cross sectional area was determined on H&E stained sections as adapted from the methods described by Frustaci and colleagues.

Renal Structure

Glomerulosclerosis Index

A minimum of 50 glomeruli were examined in PAS-stained kidney sections from each rat. The degree of sclerosis was subjectively graded on a scale of 0 to 4 as previously described: grade 0, normal; grade 1, sclerotic area up to 25% (minimal); grade 2, sclerotic area 25% to 50% (moderate); grade 3, sclerotic area 50% to 75% (moderate to severe); and grade 4, sclerotic area 75% to 100% (severe). Glomerulosclerosis was defined as basement membrane thickening, mesangial hypertrophy and capillary occlusion. A glomerulosclerosis index (GSI) was then calculated using the formula:

$\mathrm{GSI}=\sum _{i=0}^4\mathrm{Fi}\left(i\right)$
Where Fi is the percentage of glomeruli in the rat kidney with a given score (i).

Immunohistochemistry for glomerular capillary density and tubulointerstitial collagen IV Immunohistochemistry of kidney tissue was performed as previously described with antibodies in the following concentrations: JG-12 1:1000 (Bender Medsystems GbdH, Vienna, Austria) and collagen IV 1:100 (Southern Biotech). Kidney sections were scanned with the Aperio ScanScope system and analysed with ImageScope, as already described. Glomerular endothelial (JG-12) immunostaining was determined in 30 glomerular profiles from each rat kidney section. For estimation of tubulointerstitial collagen IV, the proportional area of positive immunostaining (excluding glomeruli) was determined in 10 randomly selected cortical fields (×100 magnification).

Statistics

Data are expressed as means±SEM except numerical proteinuria data which are presented as median (range). Statistical significance was determined by one-way ANOVA with a Newman-Keuls post-hoc comparison. Skew distributed urinary data were log transformed before comparison. All statistical analyses were performed using GraphPad Prism 6 for Mac OS X (GraphPad Software Inc., San Diego, Calif.). A p<0.05 was considered statistically significant.

Results

Effects of Subtotal Nephrectomy and DPP-4 Inhibition on Plasma DPP-4 Activity

In initial experiments, we set out to compare the efficacy of different doses of linagliptin and sitagliptin on plasma DPP-4 activity in sham-operated and subtotally nephectomized (SNx) rats. Since it is renally excreted, sitagliptin was “dose-adjusted” to achieve a magnitude of DPP-4 inhibition equivalent to that observed with linagliptin in SNx rats with renal impairment. Animals were therefore treated with either vehicle, 1.5 mg/kg/day linagliptin or 20 mg/kg/day sitagliptin for four days with plasma DPP-4 activity determined 1 h and 24 h after dosing. In comparison to sham-operated rats, SNx rats exhibited reduced plasma DPP-4 activity. One hour after dosing, plasma DPP-4 activity was reduced by 75-90% with either linagliptin or sitagliptin in either shamoperated or SNx rats, with no difference observed between the two inhibitors. Decreased DPP-4 activity persisted up to 24 h after dosing with linagliptin.

Effect of DPP-4 Inhibition on Metabolic Parameters in Sham and SNx Rats

Having confirmed dose-equivalency of the two DPP-4 inhibitors in SNx rats, sham and SNx rats were next randomized to receive either linagliptin 1.5 mg/kg/day, sitagliptin 20 mg/kg/day or vehicle commencing 10 days after subtotal nephrectomy or sham surgery and continuing for a further seven weeks. Sham and SNx rats treated with vehicle, linagliptin and sitagliptin. Heart weights and kidney weights were increased in SNx rats relative to sham. HbA1c was unchanged in all treatment groups. Plasma active GLP-1 levels were similarly increased by linagliptin and sitagliptin in both sham and SNx rats.

Cardiac dysfunction in subtotally nephrectomized rats

Cardiac functional and structural parameters in sham-operated and SNx rats are shown in FIGS. 1A-1G, 2A, 2B1, 2B2, 2C1, 2C2 and FIGS. 4a and 4b . The indices of cardiac dysfunction that differed significantly between vehicle-treated SNx rats and vehicle-treated sham animals are summarized in FIG. 3. This analysis revealed that SNx rats exhibited evidence of cardiac hypertrophy (increased heart weight:body weight ratio, anterior and posterior wall thickness, left ventricular mass and myocyte cross sectional area and decreased α-MHC:β-MHC ratio), cardiac fibrosis (increased collagen I mRNA and protein), diastolic dysfunction (elevated end diastolic pressure volume relationship [EDPVR]) and increased cardiac expression of both ANP and BNP (FIG. 3). In contrast, and indicative of the modest nature of cardiac dysfunction in SNx rats after eight weeks, Tau logistic and end diastolic pressure (EDP) did not differ significantly between vehicle-treated SNx and sham rats (p=0.09 and 0.12 respectively), whereas systolic function was preserved (FIGS. 4a and 4b ).

DPP-4 Inhibition Attenuates Cardiac Dysfunction in Subtotally Nephrectomized Rats

In comparison to vehicle-treated SNx rats, EDPVR was significantly lower in SNx rats treated with either linagliptin or sitagliptin (FIGS. 1A-1G). This preservation of diastolic function was accompanied by an attenuation in the overexpression of type I fibrillar collagen at both the mRNA (FIG. 2A) and protein (FIGS. 2B1 and 2B2) levels. By way of contrast, neither cardiomyocyte size determined in H&E stained sections (FIGS. 2C1 and 2C2) nor α-MHC:β-MHC ratio were changed with DPP-4 inhibition in SNx rats (α-MHC:β-MHC

ratio [AU], vehicle 0.5±0.1 [p<0.05 vs. sham], linagliptin 0.7±0.1 [p=0.16 vs. vehicle], sitagliptin 0.8±0.3 [p=0.33 vs. vehicle]).

Improved Cardiac Function with DPP-4 Inhibition Occurs Independently of Renal Change in SNx Rats

Analysis of a range of indicators of renal function and structure revealed that the beneficial effects of DPP-4 inhibition on cardiac diastolic dysfunction and fibrosis occurred independently of any change in the kidney. While, as expected, blood pressure and proteinuria were increased and GFR was reduced in SNx rats in comparison to their sham-operated counterparts, neither linagliptin nor sitagliptin significantly affected any of these parameters. Histologically, surgical renal mass ablation in SNx rats resulted in an increase in deposition of extracellular matrix within the glomeruli (reflected as an increase in glomerulosclerosis index) and cortical tubulointerstitium (illustrated by enhanced immunostaining for collagen IV). These histopathological changes were accompanied by a decrease in glomerular capillary density determined by immunostaining with the monoclonal antibody JG-12. None of these markers of renal structural injury were affected by either linagliptin or sitagliptin in either sham or SNx rats.

Neither Linagliptin Nor Sitagliptin Increase Urinary Excretion of Renal Toxicity Markers

To further exclude an improvement in renal function as a cause of the improvement in cardiac diastolic function, and to examine the “safety” of these agents when used in the context of renal impairment, we performed multi-analyte profiling of urine samples using a well-established panel of 12 renal toxicity biomarkers. Out of the screen of 12 markers, seven were increased in SNx rats in comparison to sham (B2-microglobulin, calibridin, clusterin, cystatin C, kidney injury molecule-1 [KIM-1], osteopontin and tissue inhibitor of metalloproteinases-1 [TIMP-1]), one was reduced (epidermal growth factor [EGF]), two were unaffected (glutathione-S-transferase μ [GST-μ] and vascular endothelial growth factor -A [VEGF-A]) and two were below the level of detection (GST-α and neutrophil gelatinase-associated lipocalin [NGAL]). As with the conventional markers of renal injury, none of these sensitive parameters were altered with either linagliptin or “renally adjusted” sitagliptin in either sham or SNx rats.

Claims (8)

The invention claimed is:

1. A method for treating a patient being at risk of or having heart failure with preserved ejection fraction (HFpEF), the method comprising administering to the patient 1-[(4-methyl-quinazolin-2-yl)methyl]-3-methyl-7-(2-butyn-1-yl)-8-(3-(R)-amino-piperidin-1-yl)-xanthine, optionally in combination with one or more other active agents.

2. The method of claim 1, wherein the patient has chronic kidney disease (CKD).

3. The method of claim 1, wherein the patient has severe renal impairment or end-stage renal disease (ESRD).

4. The method of claim 1, said method further comprising treating, reducing the risk of, slowing progression of, delaying the onset of, and/or protecting against heart failure with preserved ejection fraction (HFpEF) and/or diseases associated therewith.

5. The method of claim 4, wherein the disease associated with HFpEF is a major adverse cardiovascular event (MACE) selected from the group consisting of myocardial infarction, stroke, cardiovascular death, and cardiovascular hospitalization for unstable or stable angina pectoris, heart failure and coronary revascularization.

6. The method of claim 1, said method further comprising treating, reducing the risk of, slowing progression of, delaying the onset of, and/or protecting against diastolic dysfunction and/or cardiac fibrosis.

7. The method of claim 1, wherein the patient is a diabetic patient.

8. The method of claim 1, wherein the patient is a non-diabetic patient.

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